Photosynthetic Organisms Research Articles

Structure of cryptophyte photosystem II–light-harvesting antennae supercomplex

Cryptophytes are ancestral photosynthetic organisms evolved from red algae through secondary endosymbiosis. They have developed alloxanthin-chlorophyll a/c2-binding proteins (ACPs) as light-harvesting complexes (LHCs). The distinctive properties of cryptophytes contribute to efficient oxygenic photosynthesis and underscore the evolutionary relationships of red-lineage plastids. Here we present the cryo-electron microscopy structure of the Photosystem II (PSII)–ACPII supercomplex from the cryptophyte Chroomonas placoidea. The structure includes a PSII dimer and twelve ACPII monomers forming four linear trimers. These trimers structurally resemble red algae LHCs and cryptophyte ACPI trimers that associate with Photosystem I (PSI), suggesting their close evolutionary links. We also determine a Chl a-binding subunit, Psb-γ, essential for stabilizing PSII–ACPII association. Furthermore, computational calculation provides insights into the excitation energy transfer pathways. Our study lays a solid structural foundation for understanding the light-energy capture and transfer in cryptophyte PSII–ACPII, evolutionary variations in PSII–LHCII, and the origin of red-lineage LHCIIs.

The Effects of Exogenous Iron on the Photosynthetic Performance and Transcriptome of Rice under Salt–Alkali Stress

Saline-sodic stress induces iron deficiency in rice, reduces leaf photosynthetic performance, and inhibits yield enhancement. In this study, we investigated the effects of exogenous Fe on the photosynthetic performance and transcriptomics of two different tolerant rice cultivars CB9 (Changbai9: saline tolerant cultivar) and TH899 (Tonghe899: saline sensitive cultivar) with 4-week-old Fe-deficient rice seedlings under saline stress, Fe deficiency stress, and both co-stresses. The results showed that under saline and alkaline stress, spraying exogenous iron favored the growth of the two cultivars of rice, with a 32.68% and 39.82 increase in fresh weight, a 2.20-fold and 2.16-fold increase in pigment, respectively, and an 80.28% and 100.00% increase in net photosynthetic rate, respectively, as compared with the iron-deficiency treatment. Transcriptome analysis showed that we found a higher number of differentially expressed genes (7785 differentially expressed genes) in response to exogenous Fe spraying in the soda-salt sensitive variety TH899. The differentially expressed genes that are common to the two cultivars are primarily enriched in metabolic pathways, including plant hormone signal transduction (map04075) and phenylpropanoid biosynthesis (map00940). Specifically, among these genes, 14 are differentially expressed in the carotenoid biosynthetic metabolic pathway. The differentially expressed genes specific to the salinity-tolerant variety CB9 were mainly enriched in the metabolic pathways of glyoxylate and dicarboxylic acid methyl metabolism (map00630), and carbon fixation in photosynthetic organisms (map00710), among which 20 genes were significantly expressed in the pathway for carbon fixation in photosynthetic organisms (map00710). The research results offer specific theoretical support for enhancing the salt tolerance of rice.

Comparative Transcriptome-Based Analysis of the Regulation of Leaf Senescence in the Upper and Middle Canopy of Different Soybean Cultivars

Premature leaf failure is one of the major factors contributing to crop yield reduction. High-yielding soybean cultivars exhibit a longer leaf lifespan during the reproductive period, promoting higher yields. In this experiment, physiological indexes and transcriptomics analysis were carried out on leaves of different canopy parts of two soybean cultivars with different senescence sensitivities of LD32 and SND28 as test materials. The results showed that the leaf senescence rates of the two soybean cultivars, tested at the grain-filling stage, were significantly different, and the senescence rates of the upper and middle canopy leaves of LD32 were significantly lower than those of SND28. In comparison to SND28, LD32 exhibited slower decreases in chlorophyll, net photosynthetic rate, and SPAD values in the upper and middle canopy leaves. The differentially expressed genes for senescence of upper and middle canopy leaves of both cultivars were enriched in four pathways: the photosynthesis pathway, the photosynthesis–antenna protein pathway, the MAPK signaling pathway–plant hormone signal transduction pathway, and the plant hormone signal transduction pathway. The differential expression of 20 genes (Ribose-5-phosphate isomerase, fructose-1,6-bisphosphatase, etc.) in the “carbon fixation in photosynthetic organisms” pathway of LD32 may be involved in the regulation of reducing the rate of leaf senescence in the middle of the canopy at the grain-filling stage of LD32. Ribose-5-phosphate isomerase and fructose-1,6-bisphosphatase in LD32 may reduce the rate of leaf senescence in the middle of the canopy during seed filling.

An integration of physiology, transcriptomics, and proteomics reveals carbon and nitrogen metabolism responses in alfalfa (Medicago sativa L.) exposed to titanium dioxide nanoparticles

Nanoparticle (NP) pollution has negative impacts and is a major global environmental problem. However, the molecular response of alfalfa (Medicago sativa L.) to titanium dioxide nanoparticles (TiO2 NPs) is limited. Herein, the dual effects of TiO2 NPs (0–1000 mg L−1) on carbon (C) and nitrogen (N) metabolisms in alfalfa were investigated. The results showed that 500 mg L−1 TiO2 NPs (Ti-500) had the highest phytotoxicity in the C/N metabolizing enzymes; and it significantly increased total soluble sugar, starch, sucrose, and sucrose‐phosphate synthase. Furthermore, obvious photosynthesis responses were found in alfalfa exposed to Ti-500. By contrast, 100 mg L−1 TiO2 NPs (Ti-100) enhanced N metabolizing enzymes. RNA-seq analyses showed 4265 and 2121 differentially expressed genes (DEGs) in Ti-100 and Ti-500, respectively. A total of 904 and 844 differentially expressed proteins (DEPs) were identified in Ti-100 and Ti-500, respectively. Through the physiological, transcriptional, and proteomic analyses, the DEGs and DEPs related to C/N metabolism, photosynthesis, chlorophyll synthesis, starch and sucrose metabolism, and C fixation in photosynthetic organisms were observed. Overall, TiO2 NPs at low doses improve photosynthesis and C/N regulation, but high doses can cause toxicity. It is valuable for the safe application of NPs in agriculture.

Control of cyanobacterial growth with potassium; implications for bloom control in nuclear storage ponds

Microbial blooms have been reported in the First Generation Magnox Storage Pond at the Sellafield Nuclear Facility. The pond is kept alkaline with NaOH to minimise fuel rod corrosion, however alkali-tolerant microbial blooms dominated by the cyanobacterium Pseudanabaena catenata are able to thrive in this hostile environment. This study assessed the impact of alternative alkali-dosing regimens (KOH versus NaOH treatment) on biomass accumulation, using a P. catenata dominated mixed culture, which is representative of the pond environment. Optical density was reduced by 40–67 % with KOH treatment over the 3-month chemostat experiment. Microbial community analysis and proteomics demonstrated that the KOH-dependent inhibition of cell growth was mostly specific to P. catenata. The addition of KOH to nuclear storage ponds may therefore help control growth of this pioneer photosynthetic organism due to its sensitivity to potassium, while maintaining the high pH needed to inhibit the corrosion of stored nuclear fuel.

Exploring the Structure and Substance Metabolism of a Medicago sativa L. Stem Base.

The stem base of alfalfa is a critical part for its overwintering, regeneration, and yield. To better understand the specificity and importance of the stem base, we analyzed the structure, metabolic substances, and transcriptome of the stem base using anatomical techniques, ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), and RNA sequencing (RNA-seq), and compared it with stems and roots. The anatomical structure shows that the ratio of xylem to phloem changes at the base of the stem. A total of 801 compounds involved in 91 metabolic pathways were identified from the broadly targeted metabolome. Transcriptome analysis revealed 4974 differentially expressed genes (DEGs) at the stem base compared to the stem, and 5503 DEGs compared to the root. Comprehensive analyses of differentially accumulated compounds (DACs) and DEGs, in the stem base vs. stem, identified 10 valuable pathways, including plant hormone signal transduction, zeatin biosynthesis, α-Linolenic acid metabolism, histidine metabolism, carbon metabolism, carbon fixation in photosynthetic organisms, pentose phosphate pathway, galactose metabolism, and fructose and mannose metabolism. The pathways of plant hormone signal transduction and carbon metabolism were also identified by comparing the stem base with the roots. Taken together, the stem base of alfalfa is the transition region between the stem and root in morphology; in terms of material metabolism, its growth, development, and function are regulated through hormones and sugars.

DYE 10-N-NONYL ACRIDINE ORANGE AS A FLUORESCENT INDICATOR OF THE ACTION OF THE ANTISEPTIC OCTENIDINE ON THE MEMBRANES OF RHODOBACTER SPHAEROIDES CHROMATOPHORES

Increasing volumes of produced antiseptics and disinfectants, which are widely used in medicine, veterinary medicine, the food industry and everyday life, can create a serious environmental problem after use. Previously, under the infl uence of micromolar concentrations of a number of antiseptics, we identifi ed disturbances in the functioning of photosynthetic membranes and phototransforming pigment-protein complexes isolated from them in various representatives of photosynthetic organisms. In this work, to determine the sensitivity of photosynthetic membranes to the action of the cationic antiseptic octenidine, we used chromatophores of the purple nonsulfur bacteria Rhodobacter sphaeroides, labeled with the fluorescent dye 10-N-nonylacridine orange (NAO). It was shown that the binding of NAO to chromatophores is accompanied by a shift in the dye emission maximum from 525 to 640 nm. The “red” fluorescence of NAO associated with chromatophores turned out to be sensitive to the effect of increasing concentrations of octenidine on photosynthetic membranes. We have found antiseptic concentrations which led to the degradation of chromatophore structures and change in the NAO aggregative state. It can be detected by an increase of “green” fluorescence in the emission spectra of the dye. The properties of NAO as a fluorescent indicator of the functional state of photosynthetic membranes and possible changes that can occur in such systems under the influence of a cationic antiseptic are discussed.

Insightful analysis of cyanobacterial-assisted degradation of different polymers and their kinetics

The usage of plastics has increased reliably over the years. Worldwide plastic pollution has grown to 460 million tons annually, increasing global greenhouse gas emissions by 3.4 %. In recent years, several studies have been performed utilizing microorganisms for bioremediation. The aggregation of microplastics affects marine aquatic life, posing a severe threat to the aquatic environment and hurting the global food chain. Bioremediation of various polymers is cost-effective and environmentally feasible compared to chemical, physical, and biophysical methods. Cyanobacteria, also known as blue-green algae, are photosynthetic organisms and can generate value-added products from waste, i.e., waste valorization and minimization. The present study uses Cyanobacterial sp. (Anabaena & Nostoc) to degrade the synthetic polymers prepared in lab-scale conditions. Weight loss of the degraded polymers was examined, and polymeric characteristics were analyzed through SEM, FTIR, XRD, and AFM. The experimental study observed that the polymers degraded by about 70 % and 82 % by Nostoc and Anabaena sp. in about eight weeks (approx. Two months). Polymers are degraded into monomers such as vinyl alcohol, ethylene, terephthalic acid, polycaprolactone from polyethylene terephthalate, and polyvinyl alcohol. They further break down into carbon dioxide, hydrogen, and water. These are identified from the LCMS-MS analysis. The experimental data for the degradation of polymers in Cyanobacteria sp. and control media fitted the Sinclair and Peppas kinetic model well. The results inferred that those cyanobacteria could degrade the polymers very effectively since it is a potent bioremediatory agent that can remove different organic and inorganic contaminants from the environment.

Comparative physiological and transcriptomic characterization of rice (Oryza Sativa L.) root seedlings under phosphorus deficiency

Phosphorus (P) is essential for optimal growth, development, and metabolic functioning in rice. In this research study, we compared the physiologies and transcriptomes of root seedlings of two rice varieties, upland rice (LH1) and lowland rice (IR26), under the treatments of normal P application (NP: 300 μM) and low P application (LP: 10 μM). The results demonstrated that the plant biomass of LH1 and IR26 was reduced by 10.0% and 27.8% in the LP treatment compared to the NP treatment (control), respectively. The P deficiency substantially affected the enzymatic activities in the roots of LH1 and IR26, which are vital for carbon and energy metabolism. Transcriptomic results identified 24,193 genes in all samples, and over 5000 differentially expressed genes (DEGs) were identified in both varieties under LP compared with their controls. KEGG analysis showed the functional categories for DEGs, which mainly included phenylpropanoid biosynthesis, glutathione metabolism, plant hormone signal transduction, photosynthesis, and carbon fixation in photosynthetic organisms. The DEGs involved in the glutathione metabolism, carbon fixation, and photosynthesis in photosynthetic pathways were downregulated by P deficiency. The DEGs involved in the metabolism of plant hormone signal transduction and carbohydrate transition for LH1 and IR26 exhibited different defense responses to the P deficiency in upland and lowland rice. The genes engaged in plant hormone signal transduction and carbohydrate fixation are anticipated to play key roles in improving the upland and lowland rice adaptation to P-deficient conditions. The results of this investigation have established a theoretical basis for strengthening our insight into the P tolerance mechanism in lowland and upland rice.

The activation of Chlamydomonas reinhardtii alpha amylase 2 by glutamine requires its N-terminal aspartate kinase-chorismate mutase-tyrA (ACT) domain.

The coordination of assimilation pathways for all the elements that make up cellular components is a vital task for every organism. Integrating the assimilation and use of carbon (C) and nitrogen (N) is of particular importance because of the high cellular abundance of these elements. Starch is one of the most important storage polymers of photosynthetic organisms, and a complex regulatory network ensures that biosynthesis and degradation of starch are coordinated with photosynthetic activity and growth. Here, we analyzed three starch metabolism enzymes of Chlamydomonas reinhardtii that we captured by a cyclic guanosine monophosphate (cGMP) affinity chromatography approach, namely, soluble starch synthase STA3, starch-branching enzyme SBE1, and α-amylase AMA2. While none of the recombinant enzymes was directly affected by the presence of cGMP or other nucleotides, suggesting an indirect binding to cGMP, AMA2 activity was stimulated in the presence of L-glutamine (Gln). This activating effect required the enzyme's N-terminal aspartate kinase-chorismate mutase-tyrA domain. Gln is the first N assimilation product and not only a central compound for the biosynthesis of N-containing molecules but also a recognized signaling molecule for the N status. Our observation suggests that AMA2 might be a means to coordinate N and C metabolism at the enzymatic level, increasing the liberation of C skeletons from starch when high Gln levels signal an abundance of assimilated N.

Chilling stress response in tobacco seedlings: insights from transcriptome, proteome, and phosphoproteome analyses.

Tobacco (Nicotiana tabacum L.) is an important industrial crop, which is sensitive to chilling stress. Tobacco seedlings that have been subjected to chilling stress readily flower early, which seriously affects the yield and quality of their leaves. Currently, there has been progress in elucidating the molecular mechanisms by which tobacco responds to chilling stress. However, little is known about the phosphorylation that is mediated by chilling. In this study, the transcriptome, proteome and phosphoproteome were analyzed to elucidate the mechanisms of the responses of tobacco shoot and root to chilling stress (4 °C for 24 h). A total of 6,113 differentially expressed genes (DEGs), 153 differentially expressed proteins (DEPs) and 345 differential phosphopeptides were identified in the shoot, and the corresponding numbers in the root were 6,394, 212 and 404, respectively. This study showed that the tobacco seedlings to 24 h of chilling stress primarily responded to this phenomenon by altering their levels of phosphopeptide abundance. Kyoto Encyclopedia of Genes and Genomes analyses revealed that starch and sucrose metabolism and endocytosis were the common pathways in the shoot and root at these levels. In addition, the differential phosphopeptide corresponding proteins were also significantly enriched in the pathways of photosynthesis-antenna proteins and carbon fixation in photosynthetic organisms in the shoot and arginine and proline metabolism, peroxisome and RNA transport in the root. These results suggest that phosphoproteins in these pathways play important roles in the response to chilling stress. Moreover, kinases and transcription factors (TFs) that respond to chilling at the levels of phosphorylation are also crucial for resistance to chilling in tobacco seedlings. The phosphorylation or dephosphorylation of kinases, such as CDPKs and RLKs; and TFs, including VIP1-like, ABI5-like protein 2, TCP7-like, WRKY 6-like, MYC2-like and CAMTA7 among others, may play essential roles in the transduction of tobacco chilling signal and the transcriptional regulation of the genes that respond to chilling stress. Taken together, these findings provide new insights into the molecular mechanisms and regulatory networks of the responses of tobacco to chilling stress.

Widefield Super-Resolution Infrared Spectroscopy and Imaging of Autofluorescent Biological Materials and Photosynthetic Microorganisms Using Fluorescence Detected Photothermal Infrared (FL-PTIR).

We have demonstrated high-speed, super-resolution infrared (IR) spectroscopy and chemical imaging of autofluorescent biomaterials and organisms using camera-based widefield photothermal detection that takes advantage of temperature-dependent modulations of autofluorescent emission. A variety of biological materials and photosynthetic organisms exhibit strong autofluorescence emission under ultraviolet excitation and the autofluorescent emission has a very strong temperature dependence, of order 1%/K. Illuminating a sample with pulses of IR light from a wavelength-tunable laser source causes periodic localized sample temperature increases that result in a corresponding transient decrease in autofluorescent emission. A low-cost light-emitting diode-based fluorescence excitation source was used in combination with a conventional fluorescence microscopy camera to detect localized variations in autofluorescent emission over a wide area as an indicator of localized IR absorption. IR absorption image stacks were acquired over a range of IR wavelengths, including the fingerprint spectral range, enabling extraction of localized IR absorption spectra. We have applied widefield fluorescence detected photothermal IR (FL-PTIR) to an analysis of autofluorescent biological materials including collagen, leaf tissue, and photosynthetic organisms including diatoms and green microalgae cells. We have also demonstrated the FL-PTIR on live microalgae in water, demonstrating the potential for label-free dynamic chemical imaging of autofluorescent cells.

Ultrafast energy quenching mechanism of LHCSR3-dependent photoprotection in Chlamydomonas

Photosynthetic organisms have evolved an essential energy-dependent quenching (qE) mechanism to avoid any lethal damages caused by high light. While the triggering mechanism of qE has been well addressed, candidates for quenchers are often debated. This lack of understanding is because of the tremendous difficulty in measuring intact cells using transient absorption techniques. Here, we have conducted femtosecond pump-probe measurements to characterize this photophysical reaction using micro-sized cell fractions of the green alga Chlamydomonas reinhardtii that retain physiological qE function. Combined with kinetic modeling, we have demonstrated the presence of an ultrafast excitation energy transfer (EET) pathway from Chlorophyll a (Chl a) Qy to a carotenoid (car) S1 state, therefore proposing that this carotenoid, likely lutein1, is the quencher. This work has provided an easy-to-prepare qE active thylakoid membrane system for advanced spectroscopic studies and demonstrated that the energy dissipation pathway of qE is evolutionarily conserved from green algae to land plants.

Direct interaction between marine cyanobacteria mediated by nanotubes.

Microbial associations and interactions drive and regulate nutrient fluxes in the ocean. However, physical contact between cells of marine cyanobacteria has not been studied thus far. Here, we show a mechanism of direct interaction between the marine cyanobacteria Prochlorococcus and Synechococcus, the intercellular membrane nanotubes. We present evidence of inter- and intra-genus exchange of cytoplasmic material between neighboring and distant cells of cyanobacteria mediated by nanotubes. We visualized and measured these structures in xenic and axenic cultures and in natural samples. We show that nanotubes are produced between living cells, suggesting that this is a relevant system of exchange material in vivo. The discovery of nanotubes acting as exchange bridges in the most abundant photosynthetic organisms in the ocean may have important implications for their interactions with other organisms and their population dynamics.

Photosynthetic cultivation of Chlamydomonas reinhardtii with formate as a novel carbon source to the protein production

Formate, which can be produced from CO2 with green energy, has received increasing attentions as a carbon source for bioconversion. Microalgae have the native advantages of use captured light energy to facilitate the formate assimilation. However, the low efficient natural conversion and inhibition to the electron transfer chain obstacle its applications’ development. Here, using green microalga C. reinhardtii 21gr as the model, an approach to improve formate-based biomass and protein production was raised and verified through isotope labeling experiments, and the pathway of formate which presented in photosynthetic organisms was related to amino acids. By the enhancement of chloroplast glyceraldehyde-3-phosphate dehydrogenase (cGAPDH) expression, together with the adaptive laboratory evolution (ALE) over regular lethal dose of formate, the yields of the biomass and proteins were promoted with 34 % and 55 % to the wild type, respectively. Indicated by the photosynthetic activity variations, it was proposed that the balance of cellular energy and reduction power is necessary for microalgae to achieve high productivity with formate. Further investigated by proteomics, 6-phosphogluconate dehydrogenase (6-PGD) was identified as the key enzyme to formate and nitrogen’s co-assimilation.

Structure, mechanism, and evolution of the last step in vitamin C biosynthesis

Photosynthetic organisms, fungi, and animals comprise distinct pathways for vitamin C biosynthesis. Besides this diversity, the final biosynthetic step consistently involves an oxidation reaction carried out by the aldonolactone oxidoreductases. Here, we study the origin and evolution of the diversified activities and substrate preferences featured by these flavoenzymes using molecular phylogeny, kinetics, mutagenesis, and crystallographic experiments. We find clear evidence that they share a common ancestor. A flavin-interacting amino acid modulates the reactivity with the electron acceptors, including oxygen, and determines whether an enzyme functions as an oxidase or a dehydrogenase. We show that a few side chains in the catalytic cavity impart the reaction stereoselectivity. Ancestral sequence reconstruction outlines how these critical positions were affixed to specific amino acids along the evolution of the major eukaryotic clades. During Eukarya evolution, the aldonolactone oxidoreductases adapted to the varying metabolic demands while retaining their overarching vitamin C-generating function.

Photosynthetıc Energy Nanogenerators

Photosynthetıc Energy Nanogenerators

Effect of cold stress on photosynthetic physiological characteristics and molecular mechanism analysis in cold-resistant cotton (ZM36) seedlings.

Low temperature and cold damage seriously hinder the growth, development, and morphogenesis of cotton seedlings. However, the response mechanism of cotton seedlings under cold stress still lacks research. In this study, transcriptome sequencing, gas exchange parameters, and rapid chlorophyll fluorescence parameters were analyzed in leaves of cold-tolerant upland cotton variety "ZM36" under different temperature stress [25°C (T25, CK), 15°C (T15), 10°C (T10), and 4°C (T4)]. The results showed that the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), PSII potential maximum photochemical efficiency (Fv/Fm), and performance index (PIabs) of cotton leaves significantly decreased, and the intercellular CO2 concentration (Ci) and Fo/Fm significantly increased under cold stress. The transcriptome sequencing analysis showed that a total of 13,183 DEGs were involved in the response of cotton seedlings at each temperature point (T25, T15, T10, and T4), mainly involving five metabolic pathways-the phosphatidylinositol signaling system, photosynthesis, photosynthesis antenna protein, carbon fixation in photosynthetic organisms, and carotenoid synthesis. The 1,119 transcription factors were discovered among all the DEGs. These transcription factors involve 59 families, of which 15.8% of genes in the NAC family are upregulated. Through network regulatory analysis, the five candidate genes GhUVR8 (GH_A05G3668), GhPLATZ (GH_A09G2161), GhFAD4-1 (GH_A01G0758), GhNFYA1 (GH_A02G1336), and GhFAD4-2 (GH_D01G0766) were identified in response to cold stress. Furthermore, suppressing the expression level of GhPLATZ by virus-induced gene silencing led to the reduction of low temperature resistance, implying GhPLATZ as a positive regulator of low temperature tolerance. The findings of the study revealed a piece of the complex response mechanism of the cold-tolerant variety "ZM36" to different cold stresses and excavated key candidate genes for low temperature response, which provided support for accelerating the selection and breeding of cotton varieties with low temperature tolerance.

Investigating the Balance between Structural Conservation and Functional Flexibility in Photosystem I.

Photosynthesis, as the primary source of energy for all life forms, plays a crucial role in maintaining the global balance of energy, entropy, and enthalpy in living organisms. Among its various building blocks, photosystem I (PSI) is responsible for light-driven electron transfer, crucial for generating cellular reducing power. PSI acts as a light-driven plastocyanin-ferredoxin oxidoreductase and is situated in the thylakoid membranes of cyanobacteria and the chloroplasts of eukaryotic photosynthetic organisms. Comprehending the structure and function of the photosynthetic machinery is essential for understanding its mode of action. New insights are offered into the structure and function of PSI and its associated light-harvesting proteins, with a specific focus on the remarkable structural conservation of the core complex and high plasticity of the peripheral light-harvesting complexes.

Antisense RNA regulates glutamine synthetase in a heterocyst-forming cyanobacterium.

Glutamine synthetase (GS) is a key enzyme involved in nitrogen assimilation and the maintenance of C/N balance, and it is strictly regulated in all bacteria. In cyanobacteria, GS expression is controlled by nitrogen control A (NtcA) transcription factor, which operates global nitrogen regulation in these photosynthetic organisms. Furthermore, posttranslational regulation of GS is operated by protein-protein interaction with GS inactivating factors (IFs). In this study, we describe an additional regulatory mechanism involving an antisense RNA. In Nostoc sp. PCC 7120, the gifA gene (encoding GS inactivating factor IF7) is transcribed downstream of the GS (glnA) gene, from the opposite strand, and the gifA mRNA extends into the glnA coding sequence in antisense orientation. Therefore, the dual RNA transcript that encodes gifA constitutes two functional regions: a 5' protein-coding region, encoding IF7, and a 3' untranslated region that acts as an antisense to glnA. By increasing the levels of such antisense RNA either in cis or in trans, we demonstrate that the amount of GS activity can be modulated by the presence of the antisense RNA. The tail-to-tail disposition of the glnA and gifA genes observed in many cyanobacterial strains from the Nostocales clade suggests the prevalence of such antisense RNA-mediated regulation of GS in this group of cyanobacteria.